Difference between revisions of "Team:NCKU Tainan/Notebook Functional Test"

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   </head>
 
   </head>
 
   <body>
 
   <body>
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     <style>@font-face {  font-family: 'NotoSansCJKtc-Regular';  src: url("/wiki/images/0/0b/T--NCKU_Tainan--NotoSansCJKtc-Regular.woff") format('woff');}</style><nav class="navbar navbar-default"><div class="container-fluid" style="width:72%">  <div class="navbar-header">      <button type="button" class="navbar-toggle collapsed" data-toggle="collapse" data-target="#navbar" aria-expanded="false">        <span class="sr-only">Toggle navigation</span>        <span class="icon-bar"></span>        <span class="icon-bar"></span>        <span class="icon-bar"></span>      </button>      <a class="navbar-brand" href="/Team:NCKU_Tainan">        <h1>NCKU</h1><h4>Tainan</h4>      </a>  </div>  <div id="navbar" class="navbar-collapse collapse">    <ul class="nav navbar-nav">      <li>        <a class="dropdown-toggle" type="button" id="dropdownMenu1" data-toggle="dropdown" aria-haspopup="true" aria-expa="" nded="true" href="">Project</a><div class="nav-underline"></div>        <ul class="dropdown-menu" aria-labelledby="dropdownMenu2">          <li><a href="/Team:NCKU_Tainan/Project">Background</a></li>          <li><a href="/Team:NCKU_Tainan/Description">Description</a></li>          <li><a href="/Team:NCKU_Tainan/Results">Results</a></li>          <li><a href="/Team:NCKU_Tainan/Model">Modeling</a></li>          <li><a href="/Team:NCKU_Tainan/Parts">Parts</a></li>        </ul>      </li>      <li>        <a class="dropdown-toggle" type="button" id="dropdownMenu2" data-toggle="dropdown" aria-haspopup="true" aria-expa="" nded="true" href="">Device</a><div class="nav-underline"></div>        <ul class="dropdown-menu" aria-labelledby="dropdownMenu2">          <li><a href="/Team:NCKU_Tainan/Hardware">Hardware</a></li>          <li><a href="/Team:NCKU_Tainan/Software">Software</a></li>          <li><a href="/Team:NCKU_Tainan/Demonstrate">Demonstrate</a></li>        </ul>      </li>      <li>        <a class="dropdown-toggle" type="button" id="dropdownMenu3" data-toggle="dropdown" aria-haspopup="true" aria-expa="" nded="true" href="">Judging</a><div class="nav-underline"></div>        <ul class="dropdown-menu" aria-labelledby="dropdownMenu3">          <li><a href="/Team:NCKU_Tainan/Medal">Medal</a></li>          <li><a href="/Team:NCKU_Tainan/Safety">Safety</a></li>        </ul>        </li>      <li>        <a class="dropdown-toggle" type="button" id="dropdownMenu4" data-toggle="dropdown" aria-haspopup="true" aria-expa="" nded="true" href="/Team:NCKU_Tainan/Team">Team</a><div class="nav-underline"></div>        <ul class="dropdown-menu" aria-labelledby="dropdownMenu4">          <li><a href="/Team:NCKU_Tainan/Team">Team</a></li>          <li><a href="/Team:NCKU_Tainan/Attributions">Attributions</a></li>          <li><a href="/Team:NCKU_Tainan/Acknowledgement">Acknowledgement</a></li>          <li><a href="/Team:NCKU_Tainan/Collaborations">Collaborations</a></li>        </ul>        </li>      <li>        <a class="dropdown-toggle" type="button" id="dropdownMenu5" data-toggle="dropdown" aria-haspopup="true" aria-expa="" nded="true" href="/Team:NCKU_Tainan/Human_Practices">Human Practices</a><div class="nav-underline"></div>        <ul class="dropdown-menu" aria-labelledby="dropdownMenu5">          <li><a href="/Team:NCKU_Tainan/Human_Practices">Overview</a></li>          <li><a href="/Team:NCKU_Tainan/Integrated_Practices">Integrated Practices</a></li>          <li><a href="/Team:NCKU_Tainan/Engagement">Engagement</a></li>        </ul>      </li>      <li>        <a class="dropdown-toggle" type="button" id="dropdownMenu6" data-toggle="dropdown" aria-haspopup="true" aria-expa="" nded="true" href="">Notebook</a><div class="nav-underline"></div>        <ul class="dropdown-menu" aria-labelledby="dropdownMenu6">          <li><a href="/Team:NCKU_Tainan/Notebook_Construction">Construction</a></li>          <li><a href="/Team:NCKU_Tainan/Notebook_Functional_Test">Functional Test</a></li>          <li><a href="/Team:NCKU_Tainan/Notebook_Device_Design">Device Design</a></li>          <li><a href="/Team:NCKU_Tainan/Notebook_Protocols">Protocols</a></li>          <li><a href="/Team:NCKU_Tainan/Notebook_Model">Model</a></li>        </ul>      </li>    </ul>    <ul class="nav navbar-nav navbar-right">    </ul>  </div><!--/.nav-collapse --></div><!--/.container-fluid --></nav><div id="container-big"><div id="iGEMbar"></div><div id="line-left"></div><div id="line-left2"></div><div id="line-right"></div><div id="photo-left"></div></div><!--/.container-big -->        <style>
+
     <style>@font-face {  font-family: 'NotoSansCJKtc-Regular';  src: url("/wiki/images/0/0b/T--NCKU_Tainan--NotoSansCJKtc-Regular.woff") format('woff');}</style><nav class="navbar navbar-default"><div id="navbar-container" class="container-fluid">  <div class="navbar-header">      <button type="button" class="navbar-toggle collapsed" data-toggle="collapse" data-target="#navbar" aria-expanded="false">        <span class="sr-only">Toggle navigation</span>        <span class="icon-bar"></span>        <span class="icon-bar"></span>        <span class="icon-bar"></span>      </button>      <a class="navbar-brand" href="/Team:NCKU_Tainan">        <h1>NCKU</h1><h4>Tainan</h4>      </a>  </div>  <div id="navbar" class="navbar-collapse collapse">    <ul class="nav navbar-nav">      <li>        <a class="dropdown-toggle" type="button" id="dropdownMenu1" data-toggle="dropdown" aria-haspopup="true" aria-expa="" nded="true" href="">Project</a><div class="nav-underline"></div>        <ul class="dropdown-menu" aria-labelledby="dropdownMenu2">          <li><a href="/Team:NCKU_Tainan/Project">Background</a></li>          <li><a href="/Team:NCKU_Tainan/Description">Description</a></li>          <li><a href="/Team:NCKU_Tainan/Results">Results</a></li>          <li><a href="/Team:NCKU_Tainan/Model">Modeling</a></li>          <li><a href="/Team:NCKU_Tainan/Parts">Parts</a></li>        </ul>      </li>      <li>        <a class="dropdown-toggle" type="button" id="dropdownMenu2" data-toggle="dropdown" aria-haspopup="true" aria-expa="" nded="true" href="">Device</a><div class="nav-underline"></div>        <ul class="dropdown-menu" aria-labelledby="dropdownMenu2">          <li><a href="/Team:NCKU_Tainan/Hardware">Hardware</a></li>          <li><a href="/Team:NCKU_Tainan/Software">Software</a></li>          <li><a href="/Team:NCKU_Tainan/Demonstrate">Demonstrate</a></li>        </ul>      </li>      <li>        <a class="dropdown-toggle" type="button" id="dropdownMenu3" data-toggle="dropdown" aria-haspopup="true" aria-expa="" nded="true" href="">Judging</a><div class="nav-underline"></div>        <ul class="dropdown-menu" aria-labelledby="dropdownMenu3">          <li><a href="/Team:NCKU_Tainan/Medal">Medal</a></li>          <li><a href="/Team:NCKU_Tainan/Safety">Safety</a></li>        </ul>        </li>      <li>        <a class="dropdown-toggle" type="button" id="dropdownMenu4" data-toggle="dropdown" aria-haspopup="true" aria-expa="" nded="true" href="/Team:NCKU_Tainan/Team">Team</a><div class="nav-underline"></div>        <ul class="dropdown-menu" aria-labelledby="dropdownMenu4">          <li><a href="/Team:NCKU_Tainan/Team">Team</a></li>          <li><a href="/Team:NCKU_Tainan/Attributions">Attributions</a></li>          <li><a href="/Team:NCKU_Tainan/Acknowledgement">Acknowledgement</a></li>          <li><a href="/Team:NCKU_Tainan/Collaborations">Collaborations</a></li>        </ul>        </li>      <li>        <a class="dropdown-toggle" type="button" id="dropdownMenu5" data-toggle="dropdown" aria-haspopup="true" aria-expa="" nded="true" href="/Team:NCKU_Tainan/Human_Practices">Human Practices</a><div class="nav-underline"></div>        <ul class="dropdown-menu" aria-labelledby="dropdownMenu5">          <li><a href="/Team:NCKU_Tainan/Human_Practices">Overview</a></li>          <li><a href="/Team:NCKU_Tainan/Integrated_Practices">Integrated Practices</a></li>          <li><a href="/Team:NCKU_Tainan/Engagement">Engagement</a></li>        </ul>      </li>      <li>        <a class="dropdown-toggle" type="button" id="dropdownMenu6" data-toggle="dropdown" aria-haspopup="true" aria-expa="" nded="true" href="">Notebook</a><div class="nav-underline"></div>        <ul class="dropdown-menu" aria-labelledby="dropdownMenu6">          <li><a href="/Team:NCKU_Tainan/Notebook_Construction">Construction</a></li>          <li><a href="/Team:NCKU_Tainan/Notebook_Functional_Test">Functional Test</a></li>          <li><a href="/Team:NCKU_Tainan/Notebook_Device_Design">Device Design</a></li>          <li><a href="/Team:NCKU_Tainan/Notebook_Protocols">Protocols</a></li>          <li><a href="/Team:NCKU_Tainan/Notebook_Model">Model</a></li>        </ul>      </li>    </ul>    <ul class="nav navbar-nav navbar-right">    </ul>  </div><!--/.nav-collapse --></div><!--/.container-fluid --></nav><div id="container-big"><div id="iGEMbar"></div><div id="line-left"></div><div id="line-left2"></div><div id="line-right"></div><div id="photo-left"></div></div><!--/.container-big -->        <style>
 
             #photo-left { background-image: url("/wiki/images/c/cb/T--NCKU_Tainan--Functional.png"); }
 
             #photo-left { background-image: url("/wiki/images/c/cb/T--NCKU_Tainan--Functional.png"); }
 
         </style>
 
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                         <div class="text-content">
 
                         <div class="text-content">
 
                           <h5>May 14/2016</h5>
 
                           <h5>May 14/2016</h5>
−
                           <p>Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-Pcar-RBS-amilCP-TT and pMD19T-PI-RBS-amilCP-TT from LB agar plate .</p>
+
                           <p>Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-Pcar-RBS-amilCP-TT and pMD19T-PI-RBS-amilCP-TT from LB agar plate.</p>
 
                           <h5>May 15/2016</h5>
 
                           <h5>May 15/2016</h5>
 
                           <p>Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.</p>
 
                           <p>Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.</p>
 
                           <h5>May 16/2016</h5>
 
                           <h5>May 16/2016</h5>
−
                           <p>Induction : add 100 μl of bacteria culture to 96 well plate and add glucose solution to final concentration : 0, 5, 15, 30, 60, 120 mM</p>
+
                           <p>Induction: add 100 μl of bacteria culture to 96 well plate and add glucose solution to final concentration: 0, 5, 15, 30, 60, 120 mM</p>
 
                           <p>Readouts: measure OD 580 kinetically for 3.5 hr and compare the efficiency of Pcar and PI</p>
 
                           <p>Readouts: measure OD 580 kinetically for 3.5 hr and compare the efficiency of Pcar and PI</p>
 
                         </div>
 
                         </div>
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                         <div class="text-content">
 
                         <div class="text-content">
 
                           <h5>May 20/2016</h5>
 
                           <h5>May 20/2016</h5>
−
                           <p>Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PI-RBS-amilCP-TT from LB agar plate .</p>
+
                           <p>Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PI-RBS-amilCP-TT from LB agar plate.</p>
 
                           <h5>May 21/2016</h5>
 
                           <h5>May 21/2016</h5>
 
                           <p>Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.</p>
 
                           <p>Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.</p>
−
                           <p>Induction : add 100 μl of bacteria culture to 96 well plate and add glucose solution to final concentration : 0, 5, 15, 30, 60, 120 mM.</p>
+
                           <p>Induction: add 100 μl of bacteria culture to 96 well plate and add glucose solution to final concentration: 0, 5, 15, 30, 60, 120 mM.</p>
 
                           <p>Readouts: measure OD 580 kinetically for 3.5 hr.</p>
 
                           <p>Readouts: measure OD 580 kinetically for 3.5 hr.</p>
 
                         </div>
 
                         </div>
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                         <div class="text-content">
 
                         <div class="text-content">
 
                           <h5>Jun. 17/2016</h5>
 
                           <h5>Jun. 17/2016</h5>
−
                           <p>Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PI-RBS-amilCP-TT from LB agar plate .</p>
+
                           <p>Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PI-RBS-amilCP-TT from LB agar plate.</p>
 
                           <h5>Jun. 18/2016</h5>
 
                           <h5>Jun. 18/2016</h5>
 
                           <p>Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.</p>
 
                           <p>Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.</p>
−
                           <p>Sample preparation :collect urine sample from non-diabetic healthy individuals , and add additional glucose to final urine glucose concentration of 0,10,30,120,240 mM.</p>
+
                           <p>Sample preparation: collect urine sample from non-diabetic healthy individuals, and add additional glucose to final urine glucose concentration of 0,10,30,120,240 mM.</p>
−
                           <p>Induction : add 100 μl of bacteria culture to 96 well plate and add Sample Urine to final concentration : 0, 5, 15, 30, 60, 120 mM.</p>
+
                           <p>Induction: add 100 μl of bacteria culture to 96 well plate and add Sample Urine to final concentration: 0, 5, 15, 30, 60, 120 mM.</p>
 
                           <p>Readouts: measure OD 580 kinetically for 3.5 hr.</p>
 
                           <p>Readouts: measure OD 580 kinetically for 3.5 hr.</p>
 
                         </div>
 
                         </div>
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                           <p>Culture: 10 ml of LB (with Ampicillin) cultured with 100 μl pre-cultured bacteria.</p>
 
                           <p>Culture: 10 ml of LB (with Ampicillin) cultured with 100 μl pre-cultured bacteria.</p>
 
                           <h5>Jul. 18/2016</h5>
 
                           <h5>Jul. 18/2016</h5>
−
                           <p>Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PI-RBS-mRFP from LB agar plate .</p>
+
                           <p>Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PI-RBS-mRFP from LB agar plate.</p>
 
                           <h5>Jul. 19/2016</h5>
 
                           <h5>Jul. 19/2016</h5>
 
                           <p>Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.</p>
 
                           <p>Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.</p>
−
                           <p>Induction : add 100 μl of bacteria culture to 96 well plate and add glucose solution to final concentration : 0, 5, 15, 30, 60, 120 mM.</p>
+
                           <p>Induction: add 100 μl of bacteria culture to 96 well plate and add glucose solution to final concentration: 0, 5, 15, 30, 60, 120 mM.</p>
 
                           <p>Readouts: measure fluorescence of excitation 530 / emission 590 kinetically for 2 hr.</p>
 
                           <p>Readouts: measure fluorescence of excitation 530 / emission 590 kinetically for 2 hr.</p>
 
                         </div>
 
                         </div>
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                         <div class="text-content">
 
                         <div class="text-content">
 
                           <h5>Jul. 25/2016</h5>
 
                           <h5>Jul. 25/2016</h5>
−
                           <p>Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PI-RBS-mRFP-TT from LB agar plate .</p>
+
                           <p>Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PI-RBS-mRFP-TT from LB agar plate.</p>
 
                           <h5>Jul. 26/2016</h5>
 
                           <h5>Jul. 26/2016</h5>
 
                           <p>Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.</p>
 
                           <p>Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.</p>
−
                           <p>Induction : add 100 μl of bacteria culture to 96 well plate and add glucose solution to final concentration : 0, 5, 15, 30, 60, 120 mM.</p>
+
                           <p>Induction: add 100 μl of bacteria culture to 96 well plate and add glucose solution to final concentration: 0, 5, 15, 30, 60, 120 mM.</p>
 
                           <p>Readouts: measure fluorescence of excitation 530 / emission 590 kinetically for 12 hr.</p>
 
                           <p>Readouts: measure fluorescence of excitation 530 / emission 590 kinetically for 12 hr.</p>
 
                         </div>
 
                         </div>
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                         <div class="text-content">
 
                         <div class="text-content">
 
                           <h5>Aug. 22/2016</h5>
 
                           <h5>Aug. 22/2016</h5>
−
                           <p>Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-P<sub>BAD</sub>-RBS-lysis-TT from LB agar plate .</p>
+
                           <p>Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-P<sub>BAD</sub>-RBS-lysis-TT from LB agar plate.</p>
 
                           <h5>Aug. 23/2016</h5>
 
                           <h5>Aug. 23/2016</h5>
−
                           <p>Transfer : 20 ml of modified M9 (with Ampicilin) added with different glucose/arabinose concentration and culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 15 mins and transfer to 96 well plate.</p>
+
                           <p>Transfer: 20 ml of modified M9 (with Ampicilin) added with different glucose/arabinose concentration and culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 15 mins and transfer to 96 well plate.</p>
−
                           <p>Readouts: measure OD 600 kinetically with plate readerfor 4 hours .</p>
+
                           <p>Readouts: measure OD 600 kinetically with plate readerfor 4 hours.</p>
 
                         </div>
 
                         </div>
  
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                         <div class="text-content">
 
                         <div class="text-content">
 
                           <h5>Sep. 01/2016</h5>
 
                           <h5>Sep. 01/2016</h5>
−
                           <p>Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-P<sub>BAD</sub>-RBS-lysis-TT from LB agar plate .</p>
+
                           <p>Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-P<sub>BAD</sub>-RBS-lysis-TT from LB agar plate.</p>
 
                           <h5>Sep. 02/2016</h5>
 
                           <h5>Sep. 02/2016</h5>
−
                           <p>Transfer : 20 ml of modified M9 (with Ampicilin)added with different glucose/arabinose concentration and culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 15 mins and transfer to 96 well plate.</p>
+
                           <p>Transfer: 20 ml of modified M9 (with Ampicilin)added with different glucose/arabinose concentration and culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 15 mins and transfer to 96 well plate.</p>
−
                           <p>Readouts: measure OD 600 kinetically with plate readerfor 4 hours .</p>
+
                           <p>Readouts: measure OD 600 kinetically with plate readerfor 4 hours.</p>
 
                         </div>
 
                         </div>
  
Line 99: Line 99:
 
                         <div class="text-content">
 
                         <div class="text-content">
 
                           <h5>Sep. 05/2016</h5>
 
                           <h5>Sep. 05/2016</h5>
−
                           <p>Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PI-RBS-mRFP-TT from LB agar plate .</p>
+
                           <p>Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PI-RBS-mRFP-TT from LB agar plate.</p>
 
                           <h5>Sep. 06/2016</h5>
 
                           <h5>Sep. 06/2016</h5>
 
                           <p>Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.</p>
 
                           <p>Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.</p>
−
                           <p>Sample preparation :collect urine sample from non-diabetic healthy individuals , and add additional glucose to final urine glucose concentration of 0,10,30,120,240 mM.</p>
+
                           <p>Sample preparation: collect urine sample from non-diabetic healthy individuals, and add additional glucose to final urine glucose concentration of 0,10,30,120,240 mM.</p>
−
                           <p>Induction : add 100 ul of bacteria culture to 96 well plate and add urine sample to final concentration : 0, 5, 15, 30, 60, 120 mM</p>
+
                           <p>Induction: add 100 ul of bacteria culture to 96 well plate and add urine sample to final concentration: 0, 5, 15, 30, 60, 120 mM</p>
 
                           <p>Readouts: measure fluorescence of excitation 530 / emission 590 kinetically for 2 hr.</p>
 
                           <p>Readouts: measure fluorescence of excitation 530 / emission 590 kinetically for 2 hr.</p>
 
                         </div>
 
                         </div>
Line 111: Line 111:
 
                         <div class="text-content">
 
                         <div class="text-content">
 
                           <h5>Sep. 12/2016</h5>
 
                           <h5>Sep. 12/2016</h5>
−
                           <p>Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PI-RBS-mRFP-TT from LB agar plate .</p>
+
                           <p>Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PI-RBS-mRFP-TT from LB agar plate.</p>
 
                           <h5>Sep. 13/2016</h5>
 
                           <h5>Sep. 13/2016</h5>
 
                           <p>Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.</p>
 
                           <p>Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.</p>
−
                           <p>Sample preparation :collect urine sample from non-diabetic healthy individuals , and add additional glucose to final urine glucose concentration of 0,10,30,120,240 mM.</p>
+
                           <p>Sample preparation: collect urine sample from non-diabetic healthy individuals, and add additional glucose to final urine glucose concentration of 0,10,30,120,240 mM.</p>
−
                           <p>Induction : add 100 μl of bacteria culture to 96 well plate and add urine sample to final concentration : 0, 5, 15, 30, 60, 120 mM.</p>
+
                           <p>Induction: add 100 μl of bacteria culture to 96 well plate and add urine sample to final concentration: 0, 5, 15, 30, 60, 120 mM.</p>
 
                           <p>Readouts: measure fluorescence of excitation 530 / emission 590 kinetically for 12 hr.</p>
 
                           <p>Readouts: measure fluorescence of excitation 530 / emission 590 kinetically for 12 hr.</p>
 
                         </div>
 
                         </div>

Revision as of 16:56, 18 October 2016

Notebook Functional Test - iGEM NCKU

NOTE / Functional Test
Notebook - Functional Test
Compare promoter efficiency of Pcar with PI
May 14/2016

Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-Pcar-RBS-amilCP-TT and pMD19T-PI-RBS-amilCP-TT from LB agar plate.

May 15/2016

Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.

May 16/2016

Induction: add 100 μl of bacteria culture to 96 well plate and add glucose solution to final concentration: 0, 5, 15, 30, 60, 120 mM

Readouts: measure OD 580 kinetically for 3.5 hr and compare the efficiency of Pcar and PI

Glucose-Color detection test
May 20/2016

Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PI-RBS-amilCP-TT from LB agar plate.

May 21/2016

Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.

Induction: add 100 μl of bacteria culture to 96 well plate and add glucose solution to final concentration: 0, 5, 15, 30, 60, 120 mM.

Readouts: measure OD 580 kinetically for 3.5 hr.

Urine Glucose-Color detection test
Jun. 17/2016

Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PI-RBS-amilCP-TT from LB agar plate.

Jun. 18/2016

Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.

Sample preparation: collect urine sample from non-diabetic healthy individuals, and add additional glucose to final urine glucose concentration of 0,10,30,120,240 mM.

Induction: add 100 μl of bacteria culture to 96 well plate and add Sample Urine to final concentration: 0, 5, 15, 30, 60, 120 mM.

Readouts: measure OD 580 kinetically for 3.5 hr.

2-hour Glucose-Fluoresence detection test
Jul. 15/2016

Transformed plasmid (pMD19T-PpI-B0034-E1010) to BL21 competent cell.

Jul. 16/2016

Pre-culture: 3 ml of LB (with Ampicillin) was inoculated with one colony from the agar plate.

Jul. 17/2016

Extracted plasmid and confirmed plasmid size by gel electrophoresis.
Result: The size of pMD19T-PI-RBS-mRFP was correct.

Culture: 10 ml of LB (with Ampicillin) cultured with 100 μl pre-cultured bacteria.

Jul. 18/2016

Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PI-RBS-mRFP from LB agar plate.

Jul. 19/2016

Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.

Induction: add 100 μl of bacteria culture to 96 well plate and add glucose solution to final concentration: 0, 5, 15, 30, 60, 120 mM.

Readouts: measure fluorescence of excitation 530 / emission 590 kinetically for 2 hr.

12-hour Glucose-Fluoresence detection test
Jul. 25/2016

Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PI-RBS-mRFP-TT from LB agar plate.

Jul. 26/2016

Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.

Induction: add 100 μl of bacteria culture to 96 well plate and add glucose solution to final concentration: 0, 5, 15, 30, 60, 120 mM.

Readouts: measure fluorescence of excitation 530 / emission 590 kinetically for 12 hr.

Lysis test
Aug. 22/2016

Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PBAD-RBS-lysis-TT from LB agar plate.

Aug. 23/2016

Transfer: 20 ml of modified M9 (with Ampicilin) added with different glucose/arabinose concentration and culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 15 mins and transfer to 96 well plate.

Readouts: measure OD 600 kinetically with plate readerfor 4 hours.

Lysis test-2
Sep. 01/2016

Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PBAD-RBS-lysis-TT from LB agar plate.

Sep. 02/2016

Transfer: 20 ml of modified M9 (with Ampicilin)added with different glucose/arabinose concentration and culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 15 mins and transfer to 96 well plate.

Readouts: measure OD 600 kinetically with plate readerfor 4 hours.

2-hour Urine Glucose-Fluoresence detection test
Sep. 05/2016

Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PI-RBS-mRFP-TT from LB agar plate.

Sep. 06/2016

Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.

Sample preparation: collect urine sample from non-diabetic healthy individuals, and add additional glucose to final urine glucose concentration of 0,10,30,120,240 mM.

Induction: add 100 ul of bacteria culture to 96 well plate and add urine sample to final concentration: 0, 5, 15, 30, 60, 120 mM

Readouts: measure fluorescence of excitation 530 / emission 590 kinetically for 2 hr.

12-hour Glucose-Fluoresence detection test-1
Sep. 12/2016

Pre-culture: 5 ml of LB (with Ampicilin) was inoculated with one colony of pMD19T-PI-RBS-mRFP-TT from LB agar plate.

Sep. 13/2016

Culture: 20 ml of modified M9 (with Ampicilin) culture with 200μl pre-cultured bacteria. Grow up at 37 degree for 4 hours and test OD 600 until it reached 0.55~0.65.

Sample preparation: collect urine sample from non-diabetic healthy individuals, and add additional glucose to final urine glucose concentration of 0,10,30,120,240 mM.

Induction: add 100 μl of bacteria culture to 96 well plate and add urine sample to final concentration: 0, 5, 15, 30, 60, 120 mM.

Readouts: measure fluorescence of excitation 530 / emission 590 kinetically for 12 hr.